Kish Phillip E, Tsume Yasuhiro, Kijek Paul, Lanigan Thomas M, Hilfinger John M, Roessler Blake J
TSRL, Inc., 540 Avis Drive, Suite A, Ann Arbor, Michigan 48108, USA.
Mol Pharm. 2007 Jan-Feb;4(1):95-103. doi: 10.1021/mp060025q.
Bile acids conjugated to oligoarginine-containing peptides (BACs) form complexes with DNA based on the electrostatic interactions between negatively charged phosphate groups of the nucleic acid and the positively charged side chain guanidinium groups of the oligoarginine in the BACs. Charge neutralization of both components and subsequent increases of the net positive charge of the complex combined with the water-soluble lipophilic nature of the bile acid results in changes in the physicochemistry and biological properties of the complexes. We have examined the relationship of a series of 13 BACs on their interaction with circular plasmid DNA (pDNA). The formation of soluble, low-density and insoluble, high-density complexes was analyzed using several methods. The formation of high-density complexes was dependent on the DNA concentration, and was enhanced by increasing the BAC to pDNA charge ratio. Several of the BAC:pDNA complexes demonstrated exclusion of the DNA-intercalator Hoechst 33258 from pDNA, and were also protected from DNase activity. Several BAC conjugates interacted with pDNA to form nanometer-sized particles suitable for cell transfection in vitro. Five of the 13 BACs were transfection competent as single agents, and 11 of the 13 BACs showed enhancement of transfection in combination with DOPE containing liposomes or silica nanoparticles.
与含寡聚精氨酸肽结合的胆汁酸(BACs)基于核酸带负电荷的磷酸基团与BACs中寡聚精氨酸带正电荷的侧链胍基之间的静电相互作用与DNA形成复合物。两种成分的电荷中和以及复合物净正电荷随后的增加,再加上胆汁酸的水溶性亲脂性质,导致复合物的物理化学和生物学性质发生变化。我们研究了一系列13种BACs与环状质粒DNA(pDNA)相互作用的关系。使用多种方法分析了可溶性、低密度和不溶性、高密度复合物的形成。高密度复合物的形成取决于DNA浓度,并通过增加BAC与pDNA的电荷比而增强。几种BAC:pDNA复合物表现出将DNA嵌入剂Hoechst 33258从pDNA中排除,并且还受到DNase活性的保护。几种BAC缀合物与pDNA相互作用形成适合体外细胞转染的纳米级颗粒。13种BACs中有5种作为单一试剂具有转染能力,13种BACs中有11种与含DOPE的脂质体或二氧化硅纳米颗粒联合使用时表现出转染增强作用。